Simultaneous determination of 9 food additives in fruit and vegetable juice drinks by HPLC-DAD variable wavelength gradient elution

Han Qiuzhen 1, 2, 3, Huang Rimei 1, 2, 3, Chen Yanmin 1, 2, 3

(1.

3.

Abstract: To establish a high performance liquid chromatography-diode array detection method (HPLC-DAD) variable wavelength gradient elution for the simultaneous determination of acesulfame potassium, benzoic acid, sorbic acid, sodium saccharin, lemon yellow, amaranth, 9 kinds of food additives detection methods of carmine, sunset yellow and brilliant blue

C18 chromatographic column separation, gradient elution with methanol and 0.

The fruit is good, the result is accurate, and it is suitable for the simultaneous determination of 9 food additives in fruit and vegetable juice drinks

Keywords: HPLC-DAD; fruit and vegetable juice drinks; food additives; variable wavelength; gradient elution

Food additives refer to artificial or natural substances added to food to improve the quality and color, aroma, and taste of food, as well as for the needs of anti-corrosion, preservation and processing technology [1], which can improve and enhance the sensory indicators of food and maintain food Nutritional value, increase the variety of flowers and colors, adjust the nutritional structure, extend the shelf life, and improve the processing conditions [2]

It can be seen from the national food safety supervision and sampling inspections announced in 2020 [7-8] that the use of food additives beyond the scope and the limit still exists and there are problems with the combined use of excessive amounts

Therefore, this method intends to establish an optimal absorption wavelength for 9 food additives such as acesulfame potassium, benzoic acid, sorbic acid, sodium saccharin, lemon yellow, amaranth, carmine, sunset yellow and brilliant blue, which are commonly detected in fruit and vegetable juice beverages.

1 Materials and methods

1.

Agilent 1260 high performance liquid chromatograph, equipped with a diode array detector

Methanol (GR), ammonium acetate (AR), and other experimental water

1.

1.

Accurately weigh 0.

1.

Mix the sample thoroughly, accurately weigh 5 g of the sample into a 25 mL colorimetric tube, add about 15 mL of water, vortex to mix, sonicate for 30 min, adjust the pH to 6-7 after cooling, and dilute to 25 mL with water

1.
2.
3 Chromatographic conditions

Column: C18 (4.
6×250 mm, 5 μm); Flow rate: 1.
0 mL/min; Column temperature: 30 ℃; Injection volume: 10 μL; Mobile phase: Phase B is methanol, Phase D is 0.
02 mol/L acetic acid Ammonium solution, gradient elution, detection wavelength and elution conditions are shown in Table 1
.

2 Results and analysis

2.
1 Improvement of sample preparation method

The fruit and vegetable juice beverage matrix is ​​relatively simple and does not contain protein and fat.
Therefore, the synthetic colorant can be absorbed and eluted without polyamide powder, and the food additives can be detected without the steps of precipitating protein and removing fat.
This simplifies the pretreatment process and greatly improves the work.
Efficiency
.
According to the principle of the detection method, the final pre-treatment method is determined to be to weigh 5 g of the sample into a 25 mL colorimetric tube, add about 15 mL of water, vortex to mix, sonicate for 30 minutes, and adjust the pH to 6-7 after cooling.
Make the volume to 25 mL
.
use

Filter with 0.
45 μm water phase filter membrane and use the filtrate for later use
.

2.
2 Selection of detection wavelength and gradient elution

Using DAD to collect the 210-700 nm spectra of 9 kinds of food additive mixed standard working solutions, scan the best absorption wavelength, after repeated experiments, the final determination of the variable wavelength conditions is shown in Table 1
.
The gradient elution conditions consider the solubility of each additive in the mobile phase and the separation effect of the chromatographic column on the basis of the standard.
Using methanol and 20 mmol/L ammonium acetate as the mobile phase, the separation effect of different gradient conditions is repeatedly tested.
It is determined that the gradient elution program in Table 1 can achieve good separation of 9 food additives.
The chromatogram of the mixed standard working solution is shown in

Figure 1
.

2.
3 Linearity range, detection limit and quantification limit

The 9 kinds of food additives mixed standard working solutions were measured on the computer according to the chromatographic conditions in 1.
2.
3, and the mass concentration was used as the abscissa and the peak area as the ordinate to draw a standard curve.
The standard curve, correlation coefficient, detection limit and quantification limit are shown in the table 2
.

2.
4 The recovery rate and precision of the method

Weigh 5 g of a sample with a known content, add a standard solution to the sample, perform sample processing according to 1.
2.
2, and perform computer-based determination according to the chromatographic conditions in 1.
2.
3 above.
The results are shown in Table 3
.
The recovery rate ranges from 91.
5% to 107%, and the relative standard deviation ranges from 1.
8% to 6.
8%
.

2.
5 Determination of actual fruit and vegetable juice beverage samples

Using this experimental method, 8 samples of commercially available fruit and vegetable juice drinks were measured and analyzed, and each sample was measured twice
.
The test results are shown in Table 4
.
The results found that among the 8 samples, 6 food additives including acesulfame potassium, benzoic acid, sorbic acid, tartrazine, carmine, and sunset yellow were detected at the same time.
At the same time, acesulfame potassium, sorbic acid and lemon yellow were detected at the same time.
There is 1 sample of 5 food additives including acesulfame K, sunset yellow, and brilliant blue.
At the same time, 1 sample of 4 food additives including acesulfame potassium, sorbic acid, lemon yellow and sunset yellow was detected.
Only the samples where acesulfame was detected 1 and 3 samples only detected benzoic acid
.
According to the "National Food Safety Standard Food Additives Use Standards" (GB 2760—2014) [1] for the use of food additives, the content of additives in the detected samples meets the national standards

Request
.

3 Conclusion

In this study, an HPLC-DAD variable wavelength gradient elution was established for the simultaneous determination of 9 food additives in fruit and vegetable juice beverages: acesulfame K, benzoic acid, sorbic acid, sodium saccharin, lemon yellow, amaranth, carmine, sunset yellow and brilliant blue The detection method
.
In view of the characteristics of fruit and vegetable juice beverages, the pretreatment method was optimized, and the simultaneous determination of 9 food additives was realized by adopting variable wavelength gradient elution
.
The results show that the method has simple pretreatment, good chromatographic separation effect and accurate results.
It is suitable for 9 kinds of acesulfame potassium, benzoic acid, sorbic acid, sodium saccharin, lemon yellow, amaranth, carmine, sunset yellow and brilliant blue in fruit and vegetable juice beverages.
Simultaneous determination of food additives greatly improves work efficiency, reduces experimental costs, and provides strong support and effective guarantee for food safety supervision
.

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